Assessing equine anxiety-related parameters using an isolation test in combination with a questionnaire survey

The aim of the study was to determine parameters reflecting equine anxiety trait by comparing results obtained in a behavior test and an anxiety score assessed by familiar caretakers in response to a questionnaire. In the behavior test, horses were individually led into a novel room by their caretakers and loosely tethered to decrease excessive movement using the common cross-tying technique with two leads and breakable plastic cords. The horses initially remained with their caretaker for 2 min; the caretaker then left and the subject animal was left alone for 2 min. The latency to break the plastic cords, heart rate, the number of steps and pawings were recorded. When the horses were left alone, most parameters changed significantly and some showed sexual differences. A correlation analysis revealed that anxiety score assessed by caretakers showed a negative correlation with the latency to break the plastic cord and a positive correlation with heart rate only when horses were isolated. These two were suggested feasible parameters for assessing the anxiety trait of unfamiliar horses. Both the behavioral results and the anxiety scores also indicated that females were more anxious than males. Our results suggest that it would be a useful strategy for assessing other temperament traits as well to combine behavior tests with a questionnaire survey.     

Genetic analysis of phosphoprotein and matrix protein of rabies viruses isolated in Brazil

To investigate the genetic characteristics of phosphoprotein (P) and matrix protein (M) genes of variable rabies virus (RV) prevalent in Brazil, the authors genetically characterized the P and M genes from 30 Brazilian RV field isolates. Phylogenetic analysis based on the P and M genes revealed the presence of six RV variants that consisted primarily of three insectivorous bats, the vampire bat, dog and fox in Brazil. Specific amino acid substitutions corresponding to these phylogenetic lineages were observed, with Asp(42) and Glu(62) in the P protein found to be characteristic of Brazilian chiroptera- and carnivora-related RVs, respectively. Amino acid sequence motifs predicted to associate with a viral function in the P and M proteins were conserved among Brazilian RV variants.     

Identification and validation of quantitative trait loci associated with seed yield in soybean

In soybean [Glycine max (L.) Merrill], the genetic analysis of seed yield is important to aid in the breeding of high-yielding cultivars. Seed yield is a complex trait, and the number of quantitative trait loci (QTL) involved in seed yield is high. The aims of this study were to identify QTL associated with seed yield and validate their effects on seed yield using near-isogenic lines. The QTL analysis was conducted using a recombinant inbred line population derived from a cross between Japanese cultivars ‘Toyoharuka’ and ‘Toyomusume’, and eight seed yield-associated QTL were identified. There were significant positive correlations between seed yield and the number of favorable alleles at QTL associated with seed yield in the recombinant inbred lines for three years. The effects of qSY8-1, a QTL promoting greater seed yield, was validated in the Toyoharuka background. In a two-year yield trial, the 100-seed weight and seed yield of Toyoharuka-NIL, the near-isogenic line having the Toyomusume allele at qSY8-1, were significantly greater than those of Toyoharuka (106% and 107%, respectively) without any change for days to flowering and maturity. Our results suggest that qSY8-1 was not associated with maturity genes, and contributed to the 100-seed weight.     

Toe white line separation associated with the notch (crena marginalis) on the distal phalanges in Thoroughbred yearling horses

The shape of the white line of the hoof is closely related to the shape of the notch on the dorsal distal bearing border of the distal phalanx (P3). In this study, a radiographic survey of the P3 of both forelimbs of 163 Thoroughbred yearling horses was conducted. The correlation of the depth and width of the notch were analyzed with the toe white line separation grades (0 to 3). As a result, the toe white line separation grade increased, the depth and the width of the notch also increased significantly. Radiographic examination of the P3 of the forelimbs might be useful for deciding whether to implement hoof care to prevent onset of toe white line separation.     

Overcoming hybrid lethality in a cross between Japanese pear and apple using gamma irradiation and confirmation of hybrid status using flow cytometry and SSR markers

Although there have been some attempts to obtain intergeneric hybrids between Japanese pear (Pyrus pyrifolia Nakai) and apple (Malus × domestica Borkh.) mature hybrids have not been generated due to hybrid lethality. Shoots from immature hybrid embryos were gamma irradiated and cultured at normal temperature conditions to obtain viable intergeneric plants. Gamma irradiation doses of 50 and 100 Gy cultured at 25 °C killed all shoots but there was one survivor at 150 Gy and 25 °C. Hybrid status of the shoot was confirmed by using morphology, flow cytometry and SSR markers. The intergeneric plant had similar leaf serrations to apple and similar petioles to Japanese pear. Flow cytometric analysis revealed that the intergeneric shoot had a DNA content intermediate between the parents. SSR analysis showed one allele from each parent for all 16 SSR loci investigated. These data strongly suggest that the obtained shoot is a true hybrid between Japanese pear and apple.     

Equine nonneoplastic abnormal ovary in a draft mare with high serum anti-Müllerian hormone: a case study

We performed a standing hand-assisted laparoscopic ovariectomy in a draft mare that presented with high serum anti-Müllerian hormone (AMH) level and had an enlarged single cystic ovary. Histopathological examination revealed no tumor cell proliferation in the ovary, but the presence of a large ovarian cyst was confirmed. In the diagnosis of abnormal ovaries in mares, a comprehensive assessment should be performed, including the monitoring of ovarian morphology and biomarkers over time, to determine the disease prognosis and treatment plan. The case of this mare with a nonneoplastic abnormal ovary and increased serum AMH level was rare. We suggest that standing hand-assisted laparoscopic ovariectomy is useful for the removal of large ovaries in draft mares.     

Regeneration of protoplasts from hyphal strands ofVolvariella volvacea

A series of experiments on the preparation and regeneration of protoplasts from hyphal strands ofVolvariella volvacea (Bull. ex. Fr.) Singer were conducted with the aim of optimizing the conditions for its efficient regeneration. One commercial (Vvcl) and two wild (EAAC-0001 and EAAC-0002) strains ofV. volvacea from the Philippines were used and subjected to varying conditions to determine the most efficient means for regeneration of their protoplasts. The effects of age and type of strain, pH, type and concentration of osmotic stabilizer, enzymatic composition, treatment time, temperature, reciprocal frequency during enzymatic lysis of the cell wall, and centrifugation conditions were investigated. Results showed that the three strains ofV. volvacea had varying responses in terms of yield, size, and ability of their protoplasts to regenerate into the protoplast regeneration medium. Among the three strains, EAAC-0002 had the highest rate of regeneration. The 5-day-old culture ofV. volvacea, when subjected to a combination of 2% Novozyme 234 and 0.2% chitinase in 0.6M mannitol (pH 6.0) for 3 h at 30°C, 90 strokes/min and centrifuged at 1100 g for 10 min; produced an efficient yield of protoplasts with a relatively high regeneration rate.Part of this paper was presented at the 47th annual meeting of the Japan Wood Research Society, Kochi, Japan, April 2–5, 1997     

Phenolic constituents ofTaxus cuspidata I: lignans from the roots

The phenolic constituents of the roots ofTaxus cuspidata (Japanese yew) were investigated. Four lignans, [(+)-taxiresinol (1), (+)-lariciresinol (2), (–)-secoisolariciresinol (3), and (+)-pinoresinol (4)] were isolated and identified. The assignment of proton and carbon atoms for the lignans were finally solved by one- and twodimensional-nuclear magnetic resonance spectra. The enantiomeric excess of these lignans were determined by chiral high-performance liquid Chromatographic analyses. (+)-Lariciresinol and (–)-secoisolariciresinol were optically pure; (+)-taxiresinol was also suggested to be optically pure, although (+)-pinoresinol was not (77% enantiomeric excess).     

DDT effects on certain ATP related systems in the peripheral nervous system of the lobster Homarus americanus

Effects of DDT (1,1,1-trichloro-2,2-bis-(p-chlorophenyl) ethane) on various ATP utilizing enzymes in the lobster peripheral nerve were studied. On the basis of inhibition by ouabain and DDT, four classes of ATPase enzymes were recognized. They are: (1) ATPase activity that is sensitive to both ouabain and DDT inhibition, or Type A, (2) ATPase activity that is sensitive to DDT inhibition only, or Type B, (3) ATPase activity that is sensitive to ouabain only, and (4) ATPase activity that is not sensitive to either ouabain or DDT. The Type A ATPase is considered to be a part of the total (Na⁺K⁺) ATPase enzyme associated with the electrogenic pump. The Type B ATPase consisted of an uncharacterized Na⁺, K⁺, and Mg²⁺ stimulated ATPase and includes also a small portion of Mg²⁺ stimulated ATPase. Ca²⁺ stimulated ATPase activity was also detected but was not significantly affected by DDT. Proteins with actomyosin-like properties were also recognized to be present, though this superprecipitation process was only slightly affected by DDT.Other systems studied include the transfer of (γ-³²P) ATP to endogenous proteins and added histone in the presence and absence of c-AMP. DDT generally stimulated the process of ³²P incorporation, while it inhibited a portion of the specific c-AMP dependent protein kinase activity.It was concluded from these studies that DDT has a potential to inhibit or otherwise interfere with a variety of enzymatic reactions that utilize ATP as a substrate. Of these systems, the Type B ATPase bore overall resemblance to the possible target for DDT.